Acta Vet. Brno 2018, 87: 27-34

Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia

Andrea Radalj1, Jakov Nišavić1, Dejan Krnjaić1, Miroslav Valčić2, Tanja Jovanović3, Ljubiša Veljović4, Nenad Milić1

1University of Belgrade, Faculty of Veterinary Medicine, Department of Microbiology, Belgrade, Serbia
2University of Belgrade, Faculty of Veterinary Medicine, Department of Infectious Diseases of Animals and Diseases of Bees, Belgrade, Serbia
3University of Belgrade, Faculty of Medicine, Institute of Microbiology and Immunology, Virology Department, Belgrade, Serbia
4Scientific Veterinary Institute of Serbia, Belgrade, Serbia

Received August 27, 2017
Accepted April 3, 2018

The presence of equine herpesviruses 1, 2 and 5 (EHV-1, EHV-2 and EHV-5) was examined in 66 samples of spinal cord, submandibular lymph nodes and spleen of healthy, non-vaccinated abattoir horses from different locations in the Republic of Serbia. Virus isolation was conducted on RK-13 cell line with the confirmation of isolated viral strains by multiplex nested polymerase chain reaction. The cytopathic effect was observed 48–72 h after the first inoculation in 28 (42.4%) organ samples, and after 5 days in 11 other samples (16.7%) that were all confirmed as EHV-1. Four other samples (6.1%) that showed cytopathic effects on day 5 of the third passage were all positive for EHV-5. Additionally, EHV-1, EHV-2, and EHV-5 were directly detected in all organs by multiplex nested PCR in 46 (69.7%), 3 (4.5%), and 7 (10.6%) samples, respectively. The molecular characterization based on nucleotide sequencing of the part of the gB gene showed that Serbian EHV-1 isolates were 100% homogenous and clustered with EHV-1 strains from Turkey, the United Kingdom, the United States, and Japan. The EHV-2 strain from Serbia branched together with Turkish EHV-2 isolates with homogeneity from 96% to 98%. Serbian EHV-5 strains can be separated in one distinct cluster with isolates from Turkey and the United States with homogeneity from 98 to 99%. These data represent the first report of the molecular characterization of EHV-1, EHV-2, and EHV-5 in the horse population of the Republic of Serbia and document the first successful isolation of Serbian EHV-5 strains.


This study was realized within the Project Grant no. TR31008 financed by The Ministry of Education, Science and Technological Development of the Republic of Serbia.


17 live references